Figures S1 and S2

The FRAP technique (fluorescence recovery after photobleaching) was performed on individual COS-7 cells transfected with the mitochondrial matrix marker mito-GFP. Both panels demonstrate that after repeated photobleaching of a selected (boxed) area, mitochondria, which are fully contained in the boxed (bleached) region, lose fluorescence first as indicated by arrows. In contrast, the fluorescence of mitochondria with extensions outside of the boxed region partially recovers by “drainage” of GFP from these adjacent areas (arrowheads), demonstrating the contiguous nature of individual mitochondrial organelles. Photobleaching was performed every 5 s, and images were taken in 20 s intervals.

Figures S3 and S4

Both figures, taken from mito-GFP transfected COS-7 cells, demonstrate that YFP-BaxtailΔ184 labels the outer mitochondrial membrane (green), which circumscribes the inner membrane compartment (red MitoTracker labeling).

Supplementary data

Movies A and B are three-dimensional images that demonstrate the conversion from the worm-like to the punctiform mitochondrial phenotype in a mito-GFP transfected cell before (movie A) and during STS treatment (movie B). The effect of DrpK38A expression is demonstrated by movies D and E, showing a mito-GFP/DrpK38A cotransfected cell under identical experimental conditions before (D) and during STS treatment (E).

Supplementary data (MOV 3803 kb)

Movie C demonstrates the conversion of the worm-like into a punctiform mitochondrial phenotype, preceded by mitochondrial stretching during STS treatment (mito-GFP transfected COS-7 cell).

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